Anti cd4 clone rm4 5

Manufactured by Cytek Biosciences

The Anti-CD4 (clone RM4-5) is a fluorophore-conjugated antibody that specifically binds to the CD4 cell surface receptor. It is a tool used in flow cytometry for the identification and enumeration of CD4+ T cells.

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Lab products found in correlation

Anti p stat1, by Cell Signaling Technology (1 mentions) Anti cd8α clone 53 6, by Cytek Biosciences (1 mentions) Brefeldin a, by Thermo Fisher Scientific (1 mentions) Anti ifnγ, by Cytek Biosciences (1 mentions) Anti cd3 clone 145 2c11, by Cytek Biosciences (1 mentions) Lsrii flow cytometer, by BD (1 mentions)

2 protocols using anti cd4 clone rm4 5

Multiparametric Flow Cytometry of T Cells and Tumor Cells

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T cells were fixed for 10 min in 1.6% PFA on ice and subsequently permeabilized in 90% methanol for at least 30 min at −20 °C. Cells were stained with anti-CD4 (clone RM4–5, Tonbo) and anti-CD8α (clone 53-6.7, Tonbo). Where indicated, T cell IFNγ production was quantified using the mouse IFNγ secretion kit (Miltenyi Biotech). B16-F10 cells were fixed as above and stained with anti-pSTAT1 (58D6: Cell Signaling Technologies). Live B16-F10 cells were stained with anti-MHC-I H2-K^b (AF6-88.5: BioLegend).
Flow cytometric data were collected on an LSR II (BD Biosciences). Analysis was done with FlowJo software (TreeStar) and with custom code written in Python and available on GitHub (https://github.com/oylab/oyFlow).

Centofanti E., Wang C., Iyer S., Krichevsky O., Oyler-Yaniv A, & Oyler-Yaniv J. (2023). The spread of interferon-γ in melanomas is highly spatially confined, driving nongenetic variability in tumor cells. Proceedings of the National Academy of Sciences of the United States of America, 120(35), e2304190120.

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Isolation and Characterization of Intestinal Immune Cells

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Single-cell suspensions were prepared from the colon as previously described.³⁵ The following monoclonal antibodies were used to stain and sort cells- Epcam (clone G8.8), CD45.2 (clone 104), CD90.2 (clone 30-H12), and MHCII (clone M5/114.152). Splenic and MLN cells were processed and stimulated as previously described.³⁵ Briefly, single-cell suspensions were prepared and stimulated with phorbol-12-myristate-13-acetate and ionomycin for 4 h, with brefeldin A (00-4506, Affymetrix eBioscience) added for the last 2 h. For intracellular cytokine staining, cells were fixed and permeabilized by using a fixation and permeabilization solution (00-5523, eBioscience). Antibodies used for staining were anti-CD3 (clone 145 – 2C11), anti-CD4 (clone RM4-5), anti-CD19 (clone 1D3), anti-IFNγ (50-7311, Tonbo), and anti- TNF (506322, Biolegend). Flow cytometry data were acquired on an LSRII flow cytometer (BD Biosciences) and analyzed with FlowJo software (TreeStar).

Sharma D., Malik A., Guy C.S., Karki R., Vogel P, & Kanneganti T.D. (2017). Pyrin Inflammasome Regulates Tight Junction Integrity to Restrict Colitis and Tumorigenesis. Gastroenterology, 154(4), 948-964.e8.

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